David A. Hoagland

Microstructure of poly(vinyl alcohol) hydrogels produced by freeze/thaw cycling

To understand the reversible gelation and subsequent aging of hydrogels prepared by freeze/thaw processing of poly(vinyl alcohol) (PVOH) solutions, the microstructures of gels prepared by different freeze/thaw protocols and aged to varying extents are studied by cryogenic transmission electron microscopy, solid-state nuclear magnetic resonance, X-ray scattering, and differential scanning calorimetry (DSC). As discussed in the literature, gelation by the freeze/thaw process occurs as a homogeneous aqueous poly(vinyl alcohol) solution is cycled, perhaps multiple times, between temperatures above 0 °C and well below 0 °C. The current investigation has determined that a few percent of chain segments crystallize during the first cycle, organizing themselves into 3-8 nm primary crystallite junctions separated on an irregular mesh by an average spacing of ∼ 30 nm. Aging or imposition of additional freeze/thaw cycles augments the level of crystallinity and transforms the as-formed liquid-like microstructure, characterized in the electron microscope by rounded ∼ 30 nm pores, into a fibrillar network. Observation that the transformation occurs at fixed mesh spacing and approximately constant average crystallite size suggests the formation of secondary crystallites that do not affect network connectivity. Dendritic ice crystallization and possibly spinodal decomposition superimpose on this nanoscale structure a matrix of much larger pores.

Electric field induced instabilities at liquid/liquid interfaces

External electric fields were used to amplify thermal fluctuations at the interface between two thin liquid films. Similar to the results shown previously for the enhancement of fluctuations at the polymer/air interface, interfacial fluctuations having a well-defined wavelength were enhanced with a characteristic growth rate. A simple theoretical framework to describe the experimental observations is presented. Both experiment and model calculation show a substantial reduction in feature size as a result of the change in surface/interfacial energy when going from the thin film to the bilayer case. Experimentally, features develop nearly 50 times faster for the bilayers in comparison to the polymer/air case. These results point to a simple route by which the nanoscopic feature can be easily and rapidly produced or replicated.

Dissolution and Dissolved State of Cytochrome c in a Neat, Hydrophilic Ionic Liquid

The dissolution and dissolved molecular state of cytochrome c were investigated in the room temperature ionic liquid ethylmethylimidazolium ethylsulfate, [EMIM][EtSO4], by viscometry, optical and vibrational spectroscopies, and peroxidase activity. In dilute mixtures, viscometry demonstrated true molecular dissolution of cytochrome c in the ionic liquid and uncovered a molecular size larger than that in aqueous buffer, suggesting altered solvation or slight denaturation. The proteins heme unit absorbs light outside the spectral range masked by [EMIM], enabling conformational assessments by UV-visible and circular dichroism spectroscopies. Adding trends from fluorescence and Fourier transform infrared spectroscopy, unchanged secondary but perturbed tertiary structures were determined, consistent with the appreciable peroxidase activity measured. Different than in aqueous buffers, denaturation is not accompanied by aggregation. Results are relevant to the proposed application of ionic liquids as media for room temperature preservation of biomacromolecules.

Protein-selective coacervation with hyaluronic acid.

Selective coacervation with hyaluronic acid (HA), a biocompatible and injectable anionic polysaccharide, was used to isolate a target protein, bovine serum albumin (BSA), with 90% purity from a 1:1 mixture with a second protein of similar pI, β-lactoglobulin (BLG). This separation was attributed to the higher HA-affinity of BSA, arising from its more concentrated positive domain. The values of pH corresponding respectively to the onset of complex formation, coacervation, precipitation, and redissolution (pH(c), pHϕ, pH(p), and pH(d)) were determined as a function of ionic strength I. These pH values were related to critical values of protein charge, Z, and their dependence on I provided some insights into the mechanisms of these transitions. The higher polyanion binding affinity of BSA, deduced from its higher values of pH(c), was confirmed by isothermal titration calorimetry (ITC). Confocal laser microscopy clearly showed time-dependent coalescence of vesicular droplets into a continuous film. Comparisons with prior results for the polycation poly(diallyldimethylammonium chloride) (PDADMAC) show reversal of protein selectivity due to reversal of the polyelectrolyte charge. Stronger binding of both proteins to PDADMAC established by ITC may be related to the higher chain flexibility and effective linear charge density of this polycation.

Assembly of Acid-Functionalized Single-Walled Carbon Nanotubes at Oil/Water Interfaces

The efficient segregation of water-soluble, acid-functionalized, single-walled carbon nanotubes (SWCNTs) at the oil/water interface was induced by dissolving low-molecular-weight amine-terminated polystyrene (PS-NH2) in the oil phase. Salt-bridge interactions between carboxylic acid groups of SWCNTs and amine groups of PS drove the assembly of SWCNTs at the interface, monitored by pendant drop tensiometry and laser scanning confocal microscopy. The impact of PS end-group functionality, PS and SWCNT concentrations, and the degree of SWCNT acid modification on the interfacial activity was assessed, and a sharp drop in interfacial tension was observed above a critical SWCNT concentration. Interfacial tensions were low enough to support stable oil/water emulsions. Further experiments, including potentiometric titrations and the replacement of SWCNTs by other carboxyl-containing species, demonstrated that the interfacial tension drop reflects the loss of SWCNT charge as the pH falls near/below the intrinsic carboxyl dissociation constant; species lacking multivalent carboxylic acid groups are inactive. The trapped SWCNTs appear to be neither ordered nor oriented.

Thermoreversible Gelation of an Ionic Liquid by Crystallization of a Dissolved Polymer

Poly(ethylene glycol) (PEG) dissolves in the room-temperature ionic liquid 1-ethyl-3-methylimidazolium ethylsulfate [EMIM][EtSO(4)] above approximately 60 degrees C, the neat polymers melting temperature, and if polymer concentration and molecular weight are high enough, the solution transforms into a semitransparent gel when cooled. The modulus, reaching 100 KPa or higher, is strongly affected by PEG concentration, and self-supporting materials are made even from solutions somewhat below coil overlap. Via differential scanning calorimetry (DSC), rheology, and optical microscopy, thermoreversible solidification is traced to kinetically frustrated polymer crystallization, an established mechanism for many pairings of crystallizable polymer with aqueous or organic solvent. Optical microscopy reveals nucleation and growth of PEG crystals with a largest dimension of tens to hundreds of micrometers. Crystalline chain packing in gels is identical to that of neat PEG, and degrees of crystallization are similar. Simple preparation, nontoxicity, and vanishing volatility suggest unique new gel applications.

Polyelectrolyte Electrophoresis in a Dilute Solution of Neutral Polymers: Model Studies

Dilute neutral polymer solutions provide an effective environment for the electrophoretic separation of polyelectrolytes by molecular weight. To better understand the mechanisms responsible, this study examines how the molecular weight dependent mobility μ of poly(styrenesulfonate), termed the probe chain, depends on the molecular weight and concentration of various dilute pullulans, termed the host chains. Available for this system are nearly monodisperse probes and hosts. Carefully performed capillary electrophoresis experiments measure absolute values of μ and its distribution. Below the overlap concentration, hosts reduce μ from its free solution value in proportion to concentration. The strongest dependence of μ on probe molecular weight occurs at nearly equal probe and host chain lengths. The influence of host and probe chain lengths on μ can be understood in terms of pairwise probe−host configurational interactions. Three parametersentanglement probability, average entanglement duration, and averag...

Evolution of hierarchical structures in polyelectrolyte–micelle coacervates

We investigated the temperature-induced liquid–liquid phase separation (coacervation) of polyelectrolyte (PE)–micelle systems and the structure of the resultant coacervates. Dynamic light scattering, small angle neutron scattering and cryo-transmission electron microscopy (DLS, SANS, Cryo-TEM) were used to examine the evolution of complex structure up to the point of temperature-induced coacervation and beyond. Three diffusional modes, seen in the single phase samples and in the coexisting coacervated supernatant phases were attributed respectively to free micelles, PE–micelle complexes, and aggregates thereof. They corresponded to SANS Guinier region slopes yielding Rg ∼ 4 nm (micelles) and Rg ∼ 50 nm (unresolved complexes and aggregates). Cryo-TEM images of coacervates indicated how these subunits are organized within dense and dilute coacervate domains at larger length scales. Taken together, these results are understood to arise from the requirements of overall charge neutralization, and ion-pairing and counterion release during coacervation. We conclude that a polyelectrolyte:micelle system at incipient coacervation with charge stoichiometry ([+]/[−] > 1) donates excess polycations to other complexes in solution. In the coacervate, a similar disproportionation but at different length scales ejects excess polycations and their counterions into dilute domains. In both phases, association and desolvation are driven by counterion release, enhanced chain configurational entropy, and ion-pairing. These enthalpic and entropic forces operating in both phases could explain the structural similarities between soluble aggregates and coacervate dense domains.

Molecular weight analysis of polycations by capillary electrophoresis in a solution of neutral polymers

Under appropriate conditions, polyelectrolytes separate according to molecular weight during their electrophoretic migration through a dilute solution of inert neutral polymers. These separations facilitate a new capillary electrophoresis-based approach for high resolution and high throughput polyelectrolyte molecular weight analysis, one theoretically applicable to both polyanions and polycations. Although pioneered for DNA, a polyanion, the new method is discussed here in the context of synthetic polycations. Numerous experimental difficulties evolve from the introduction of positive solute charge, not the least of which is a strong tendency for solute adsorption on the negative capillary walls. The adsorption can be overcome by using a run buffer with a cationic surfactant that forms a dynamic yet stable positive wall coating. Feasible at nearly any pH, the surfactant approach enables robust and high-resolution polycation analysis. For illustration, we compare the separation at low pH of three protonated poly(2-vinylpyridine)s to the separation at neutral pH of the same polymers after quaternization. A good match is found. Further, electrophoretic analyses by the new method suggest how poly(2-vinylpyridine)s degrade/crosslink when exposed to quaternizing conditions for an excessive period.

A solution birefringence study of polymer elongation in spatially periodic stretching flows

The flow birefringence of dilute polymer solutions in periodically converging/diverging channels has been employed to study the dynamics of flexible chain molecules under transient stretching stresses. The onset of periodic birefringence for chains of high molecular weight is only observed after the chains have experienced several cycles of stress, at a point deep into the channel. This slow onset indicates that the solutions possess a memory on time scales much greater than that normally associated with a relaxed flexible coil. This behavior is recorded for both aqueous and nonaqueous solutions, at concentrations both above and below the entanglement concentration. Centerline birefringence, however, which is associated with purely elongational flow, is only observed for aqueous polymer solutions. An explanation for these birefringence results is suggested, based on a configuration-dependent dumbbell model for polymer chains in dilute solution.